Morpholino oligomers tested in vitro, in biofilm and in vivo against multidrug-resistant Klebsiella pneumoniae

Bruce L. Geller, Lixin Li, Fabian Martinez, Erin Sully, Carolyn R. Sturge, Seth M. Daly, Christine Pybus, David E. Greenberg

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: Klebsiella pneumoniae is an opportunistic pathogen and many strains are multidrug resistant. KPC is one of the most problematic resistance mechanisms, as it confers resistance to most β-lactams, including carbapenems. A promising platform technology for treating infections caused by MDR pathogens is the nucleic acid-like synthetic oligomers that silence bacterial gene expression by an antisense mechanism. Objectives: To test a peptide-conjugated phosphorodiamidate morpholino oligomer (PPMO) in a mouse model of K. pneumoniae infection. Methods: PPMOs were designed to target various essential genes of K. pneumoniae and screened in vitro against a panel of diverse strains. The most potent PPMOs were further tested for their bactericidal effects in broth cultures and in established biofilms. Finally, a PPMO was used to treat mice infected with a KPC-expressing strain. Results: The most potent PPMOs targeted acpP, rpmB and ftsZ and had MIC75s of 0.5, 4 and 4 lM, respectively. AcpP PPMOs were bactericidal at 1-2×MIC and reduced viable cells and biofilm mass in established biofilms. In a mouse pneumonia model, therapeutic intranasal treatment with ~30 mg/kg AcpP PPMO improved survival by 89% and reduced bacterial burden in the lung by ~3 logs. Survival was proportional to the dose of AcpP PPMO. Delaying treatment by 2, 8 or 24 h post-infection improved survival compared with control groups treated with PBS or scrambled sequence (Scr) PPMOs. Conclusions: PPMOs have the potential to be effective therapeutic agents against KPC-expressing, MDR K. pneumoniae.

Original languageEnglish (US)
Pages (from-to)1611-1619
Number of pages9
JournalJournal of Antimicrobial Chemotherapy
Volume73
Issue number6
DOIs
StatePublished - Jun 1 2018

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Morpholinos
Klebsiella pneumoniae
Biofilms
Klebsiella Infections
Bacterial Genes
Lactams
Carbapenems
Essential Genes
Infection
Nucleic Acids
Pneumonia
Technology
Gene Expression
Lung
Control Groups
Peptides
In Vitro Techniques
Therapeutics

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases

Cite this

Morpholino oligomers tested in vitro, in biofilm and in vivo against multidrug-resistant Klebsiella pneumoniae. / Geller, Bruce L.; Li, Lixin; Martinez, Fabian; Sully, Erin; Sturge, Carolyn R.; Daly, Seth M.; Pybus, Christine; Greenberg, David E.

In: Journal of Antimicrobial Chemotherapy, Vol. 73, No. 6, 01.06.2018, p. 1611-1619.

Research output: Contribution to journalArticle

Geller, Bruce L. ; Li, Lixin ; Martinez, Fabian ; Sully, Erin ; Sturge, Carolyn R. ; Daly, Seth M. ; Pybus, Christine ; Greenberg, David E. / Morpholino oligomers tested in vitro, in biofilm and in vivo against multidrug-resistant Klebsiella pneumoniae. In: Journal of Antimicrobial Chemotherapy. 2018 ; Vol. 73, No. 6. pp. 1611-1619.
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abstract = "Background: Klebsiella pneumoniae is an opportunistic pathogen and many strains are multidrug resistant. KPC is one of the most problematic resistance mechanisms, as it confers resistance to most β-lactams, including carbapenems. A promising platform technology for treating infections caused by MDR pathogens is the nucleic acid-like synthetic oligomers that silence bacterial gene expression by an antisense mechanism. Objectives: To test a peptide-conjugated phosphorodiamidate morpholino oligomer (PPMO) in a mouse model of K. pneumoniae infection. Methods: PPMOs were designed to target various essential genes of K. pneumoniae and screened in vitro against a panel of diverse strains. The most potent PPMOs were further tested for their bactericidal effects in broth cultures and in established biofilms. Finally, a PPMO was used to treat mice infected with a KPC-expressing strain. Results: The most potent PPMOs targeted acpP, rpmB and ftsZ and had MIC75s of 0.5, 4 and 4 lM, respectively. AcpP PPMOs were bactericidal at 1-2×MIC and reduced viable cells and biofilm mass in established biofilms. In a mouse pneumonia model, therapeutic intranasal treatment with ~30 mg/kg AcpP PPMO improved survival by 89{\%} and reduced bacterial burden in the lung by ~3 logs. Survival was proportional to the dose of AcpP PPMO. Delaying treatment by 2, 8 or 24 h post-infection improved survival compared with control groups treated with PBS or scrambled sequence (Scr) PPMOs. Conclusions: PPMOs have the potential to be effective therapeutic agents against KPC-expressing, MDR K. pneumoniae.",
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AU - Li, Lixin

AU - Martinez, Fabian

AU - Sully, Erin

AU - Sturge, Carolyn R.

AU - Daly, Seth M.

AU - Pybus, Christine

AU - Greenberg, David E.

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N2 - Background: Klebsiella pneumoniae is an opportunistic pathogen and many strains are multidrug resistant. KPC is one of the most problematic resistance mechanisms, as it confers resistance to most β-lactams, including carbapenems. A promising platform technology for treating infections caused by MDR pathogens is the nucleic acid-like synthetic oligomers that silence bacterial gene expression by an antisense mechanism. Objectives: To test a peptide-conjugated phosphorodiamidate morpholino oligomer (PPMO) in a mouse model of K. pneumoniae infection. Methods: PPMOs were designed to target various essential genes of K. pneumoniae and screened in vitro against a panel of diverse strains. The most potent PPMOs were further tested for their bactericidal effects in broth cultures and in established biofilms. Finally, a PPMO was used to treat mice infected with a KPC-expressing strain. Results: The most potent PPMOs targeted acpP, rpmB and ftsZ and had MIC75s of 0.5, 4 and 4 lM, respectively. AcpP PPMOs were bactericidal at 1-2×MIC and reduced viable cells and biofilm mass in established biofilms. In a mouse pneumonia model, therapeutic intranasal treatment with ~30 mg/kg AcpP PPMO improved survival by 89% and reduced bacterial burden in the lung by ~3 logs. Survival was proportional to the dose of AcpP PPMO. Delaying treatment by 2, 8 or 24 h post-infection improved survival compared with control groups treated with PBS or scrambled sequence (Scr) PPMOs. Conclusions: PPMOs have the potential to be effective therapeutic agents against KPC-expressing, MDR K. pneumoniae.

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